The sample is hydrolyzed to generate free HYP, and hydroxyproline can produce oxidation product under the action of oxidizing agent. The generated oxidation product can react with chromogenic agent to produce burgundy. The concentration of hydroxyproline can be calculated by measuring the OD value at 558 nm.
Hydroxyproline (HYP) is a derivative of proline, mainly found in collagen. Collagen is the most abundant protein in animals and makes up connective tissues such as skin, bone, cartilage, tendons and ligaments. HYP enhances the crosslinking of collagen fibers through the hydroxyl group in its structure, thus improving the stability and tensile strength of collagen. Since HYP almost only exists in collagen, its content can reflect the synthesis and catabolism of collagen. Therefore, the detection of HYP level can be used as an indicator to monitor collagen metabolism. The detection principle of this kit: the sample is hydrolyzed by the alkali reagent to generate free HYP, and the oxidation product produced under the action of the oxidant reacts with the color agent to be purple red. The substance has an absorption peak at 565 nm, and within a certain range, its absorbance is linear with the concentration. By measuring the absorbance at 565 nm, the HYP content in the sample can be calculated.
The oxidation product which produced by hydroxyproline under the action of oxidant react with dimethylaminobenzaldehyde and show a purplish red color. The content of hydroxyproline can be calculated by measuring the OD value at 550 nm.
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